ES2456504T3 - Procedures for monitoring the efficacy of anti-IL-2R antibodies in patients with multiple sclerosis - Google Patents

Abstract

A method of monitoring the efficacy of an anti-IL-2R antibody in a patient diagnosed with multiple sclerosis, comprising determining the level of HLA-DR+CD4+ T lymphocytes, wherein a decrease in the level of HLA-DR+ T lymphocytes DR+CD4+ in the patient after exposure to anti-IL-2R antibody indicates that the anti-IL-2R antibody is effective in ameliorating at least one symptom of multiple sclerosis in the treated patient.

Description

Procedures for monitoring the efficacy of anti-IL-2R antibodies in patients with multiple sclerosis

3. Background

The goal of multiple sclerosis (MS) treatment is to prevent permanent disabilities and slow the progression of the disease. A shorter term therapeutic objective is to reduce the recurrence rate. IFN-beta is the most commonly used chronic maintenance agent to treat MS. The drawbacks of IFN-beta treatment include inadequate response in at least 30% of patients after the start of IFN-beta therapy (Bertolotto, et al., 2002, J Neurosurg Psychiatry, 73 (2): 148- 153; and Durelli, 2004, J Neurol., 251 Suppl. 4: IV13-IV24), and the induction of anti-interferon antibodies in 7% to 42% of patients (Sorensen, 2003 Lancet, 362: 1184-91 ). Although other agents are used to treat MS, including corticosteroids, glatiramer acetate, mitoxantrone, and natalizumab, these agents are only partially effective in the treatment of clinical recurrences, and some carry significant safety risks.

Since the agents that are currently being used to treat MS are not completely effective in the treatment of the disease, it is desirable to identify and clinically validate markers that can be used to control the clinical response in an individual diagnosed with MS to a therapeutic agent as a medium. to optimize and / or change the treatment regime of that individual.

4. Summary

An agent that has shown promise for treating MS in clinical trials is daclizumab. Daclizumab was evaluated as a treatment for MS in a dose-based, multicenter, placebo-controlled, double-blind, randomized, phase 2 study (the CHOICE study). At the end of the 24-week dosing period, compared to the IFN-beta placebo, there was a 25% reduction in lesions that enhance the contrast with new or increased gadolinium (Gd-CEL) contrast detected by magnetic resonance imaging (MRI) ) in the 1 mg / kg group of daclizumab and a 72% reduction in the 2 mg / kg group of daclizumab (FIG. 1). Both daclizumab regimens were associated with an approximate 35% reduction in the annualized recurrence rate at 24 weeks (Montalban, X. et al., Multiple Sclerosis, 13: S 18-S 18 Suppl. 2 OCT 2007; and, Kaufman, MD, et al., Neurology, 70 (11): A220-A220 Suppl. 1 MAR 11 2008).

Blood samples were collected from patients throughout the CHOICE study and analyzed to determine the levels of immune cell subsets and associated activation markers before, during, and after treatment with daclizumab. Reversible and rapid reductions were observed in HLA-DR + CD4 + T lymphocytes, a subset of T lymphocytes, during the treatment phase with daclizumab for the dosage groups of 1 mg / kg and 2 mg / kg. Exposure-dependent reductions in the amounts of activated T lymphocytes were consistent with new or increased exposure-dependent reductions in Gd-CEL and reductions in the annualized recurrence rate.

These results indicate that changes in HLA-DR + CD4 + T lymphocyte counts can be used as a biomarker of the clinical response to daclizumab in a patient diagnosed with MS. Accordingly, the procedures described herein disclose the use of HLA-DR + CD4 + T lymphocyte counts to control the efficacy of daclizumab in a patient diagnosed with MS. In some embodiments, the method comprises determining the level of HLA-DR + CD4 + T lymphocytes in a patient diagnosed with MS after exposure to daclizumab, in which a decrease in the level of HLA-DR + CD4 + T lymphocytes indicates that daclizumab It is effective in improving at least one symptom of MS in the treated patient. Biological samples may be collected from the treated patient one or more times before, during, and / or after treatment with daclizumab.

Symptoms of MS that can be stabilized or improved using the procedures described herein include, but are not limited to, reduce the recurrence rate, establish or reduce the rate of disability progression measured by conventional values such as the Expanded Scale value. of the Disability Level (EEND), decrease the amount of new or increased Gd-CEL, and / or decrease the amount of new or increased MRI T2 lesions. The subject being treated may have relapsing forms of MS, including relapsing / remitting MS, secondary progressive MS, progressive relapsing MS, or aggravated relapsing MS. In addition to daclizumab, other antibodies against IL-2R may be used, such as monoclonal antibodies, chimeric antibodies, humanized antibodies, or fully human antibodies that specifically bind to the alpha or p55 (Tac) chain of the IL-2 receptor in the procedures. described in this document.

It should be appreciated that both the above general description and the following detailed description are exemplary and explanatory only and are not restrictive of the compositions and procedures described herein. In the present application, the use of the singular includes the plural unless specifically indicated otherwise. In addition, the use of "or" means "and / or" unless otherwise indicated. Likewise, "understand", "understand", "understand", "include," "includes" and "includes" are not intended to be limiting.

5. Brief description of the figures

FIG. 1 is a bar chart that represents the data demonstrating the reduction in lesions that enhance gadolinium contrast at the end of a 24-week dosing period with daclizumab;

FIG. 2 is a line chart that represents data demonstrating rapid and reversible reductions in activated HLA-DR + CD4 + T lymphocytes during the daclizumab treatment phase; Y

FIG. 3 is a line chart that represents the data that demonstrates the relationship between the area of the stable regimen with daclizumab under the curve (AUC) and the decreases in the counts of activated T lymphocytes HLA-DR + CD4 + in which a reduction in HLA-DR + CD4 + cells is represented as a positive number on the y axis.

6. Detailed Description

6.1 Definitions

As used herein, the following terms are intended to have the following meanings:

The term "antibody" refers to an immunoglobulin molecule that specifically binds to, or is immunologically reactive with, a particular antigen, and includes polyclonal, monoclonal, genetically engineered (e.g., rIgG) and modified antibody forms of otherwise, including but not limited to chimeric antibodies, humanized antibodies, heteroconjugate antibodies (including, for example, bispecific antibodies), antibody antigen binding fragments, including, for example, Fab ', F (ab') 2, Fab fragments, Fv, and scFv and multimeric forms of antigen-binding fragments, including, for example, diabodies, triabodies and tetrabodies. In addition, unless otherwise indicated, it is understood that the term "monoclonal antibody" (mAb) includes both intact molecules, as well as antibody fragments (such as, for example, Fab and F (ab ') 2 fragments) that They are able to specifically bind to a protein. Fab and F (ab ') 2 fragments lack the Fc fragment of the intact antibody, are removed more quickly from the animal or plant's circulation, and may have less non-specific tissue binding than an intact antibody (Wahl et al., 1983 , J. Nucl. Med. 24: 316).

The term "scFv" refers to a single chain Fv antibody in which the variable domains of the heavy chain and the light chain of a traditional antibody have joined to form a chain.

References to "VH" refer to a variable region of an immunoglobulin heavy chain of an antibody, including the heavy chain of an Fv, scFv, or Fab. References to "VL" refer to the variable region of an immunoglobulin light chain, including the light chain of an Fv, scFv, dsFv or Fab. Antibodies (Ab) and immunoglobulins (Ig) are glycoproteins that have the same structural characteristics. Although antibodies show binding specificity for a specific target, immunoglobulins include both antibodies and other antibody-like molecules that lack target specificity. Native antibodies and immunoglobulins are usually heterotetrameric glycoproteins of approximately 150,000 daltons, composed of two identical light chains (L) and two identical heavy chains (H). Each heavy chain has at one end a variable domain (VH) followed by several constant domains. Each light chain has a variable domain at one end (VL) and a constant domain at its other end.

Complementarity determining regions (CDRs) are also known as hypervariable regions in the variable domains of both the light chain and the heavy chain. The most highly conserved parts of the variable domains are called flanking regions (FR). As is known in the art, the position / limit of amino acids that delineates a hypervariable region of an antibody may vary, depending on the context and the various definitions known in the art. Some positions within a variable domain can be seen as hybrid hypervariable positions in which these positions can be considered within a hypervariable region under a series of criteria while they are considered outside a hypervariable region under a different series of criteria. One or more of these positions can also be found in extended hypervariable regions. The disclosure provides antibodies comprising modifications in these hypervariable hybrid positions. The variable domains of the native heavy and light chains each comprise four FR regions, which largely adopt a sheet configuration!, Connected by three CDRs, which form loops that connect, and in some cases are part of, the structure of sheet !. The CDRs in each chain are held together in close proximity by the FR regions and, with the CDRs of the other chain, contribute to the formation of the target binding site of the antibodies (see Kabat et al., Sequences of Proteins of Immunological Interest (National Institute of Health, Bethesda, Md. 1987) As used herein, the numbering of the immunoglobulin amino acid residues is done in accordance with the immunoglobulin amino acid residue numbering system of Kabat et al. ., unless otherwise indicated.

The term "antibody fragment" refers to a part of a full-length antibody, generally the target or variable binding region. Examples of antibody fragments include Fab, Fab ', F (ab') 2 and Fv fragments. An "Fv" fragment is the minimum antibody fragment that contains a complete recognition site and

target binding. This region consists of a dimer of a heavy chain and a light chain variable domain in a close non-covalent association (VH-VL dimer). It is in this configuration that the three CDRs of each variable domain interact to define a target binding site on the surface of the VH-VL dimer. Collectively, the six CDRs confer specificity of target binding to the antibody. However, even a single variable domain (or half of an Fv comprising only three specific CDRs for a target) has the ability to recognize and bind to the target, albeit at a lower affinity than the entire binding site. The "single chain Fv" or "scFv" antibody fragments comprise the VH and VL domains of an antibody, in which these domains are present in a single polypeptide chain. Generally, the Fv polypeptide further comprises a polypeptide linker between the VH and VL domains that allows the scFv to form the desired structure for target binding.

The Fab fragment contains the constant domain of the light chain and the first constant domain (CH1) of the heavy chain. Fab 'fragments differ from Fab fragments by the addition of a few residues at the carboxyl end of the CH1 domain of the heavy chain including one or more cysteines of the hinge region of the antibody. Fab 'fragments are produced by cleavage of the disulfide bond in the hinge cysteines of the pepsin digestion product of F (ab') 2. Those skilled in the art know additional chemical couplings of antibody fragments.

"Epitope" or "antigenic determinant" refers to a site in an antigen to which an antibody binds. Epitopes can be formed from contiguous amino acids or non-contiguous amino acids juxtaposed by tertiary folding of a protein. Epitopes formed from contiguous amino acids are typically retained on exposure to denaturing solvents while epitopes formed by tertiary folding are typically lost on treatment with denaturing solvents. An epitope typically includes at least 3, and more usually, at least 5 or 8-10 amino acids in a single spatial conformation. Methods for determining the spatial conformation of epitopes include, for example, X-ray crystallography and two-dimensional nuclear magnetic resonance. See, for example, Epitope Mapping Protocols in Methods in Molecular Biology, Vol. 66, Glenn E. Morris, Ed (1996).

The determination of whether two antibodies bind substantially to the same epitope is achieved using methods known in the art, such as a competition assay. In conducting an antibody competition study between a control antibody (eg, daclizumab) and any test antibody, the control antibody can first be labeled with a detectable label, such as biotin, an enzyme, radioactive label, or fluorescent marker to enable subsequent identification. In said assay, the intensity of the bound marker in a sample containing the labeled control antibody is measured and the intensity of the bound marker in the sample containing the control antibody and the unlabeled test antibody is measured. If the unlabeled test antibody competes with the labeled antibody by binding to an overlapping epitope, the intensity of the detected label will increase relative to the binding in the sample containing only the labeled control antibody. Other methods for determining binding are known in the art.

The term "monoclonal antibody" refers to an antibody that is obtained from a single clone, including any eukaryotic, prokaryotic, or phage clone, and not the procedure by which it is produced. Immunologically reactive antibodies with a particular antigen can be prepared using a wide variety of techniques known in the art including the use of hybridoma, recombinant, and phage display technologies,

or a combination thereof. For example, monoclonal antibodies can be produced using hybridoma techniques including those known in the art and shown, for example, in Harlow and Lane, "Antibodies: A Laboratory Manual," Cold Spring Harbor Laboratory Press, New York (1988); Hammerling et al., In: "Monoclonal Antibodies and T-Cell Hybridomas," Elsevier, N.Y. (1981), p. 563 681 (both of which are incorporated herein by reference in their entirety).

A "chimeric antibody" is an immunoglobulin molecule in which (a) the constant region, or a part thereof, is altered, replaced or exchanged so that the antigen binding site (variable region) is bound to a region constant of a different or altered class, effector function and / or species, or a completely different molecule that confers new properties to the chimeric antibody, for example, an enzyme, toxin, hormone, growth factor, drug, etc .; or (b) the variable region, or a part thereof, is altered, replaced or exchanged with a variable region that has a different or altered antigen specificity. Any of the antiIL-2R antibodies described herein may be chimeric.

The term "humanized antibody" or "humanized immunoglobulin" refers to an immunoglobulin comprising a human flanking region, at least one and preferably all complementarity determining regions (CDR) of a non-human antibody, and in which any constant region present it is substantially identical to a constant region of human immunoglobulin, that is, at least about 85%, at least 90%, and at least 95% identical. Thus, all parts of a humanized immunoglobulin, except possibly the CDRs, are substantially identical to corresponding parts of one or more native human immunoglobulin sequences. Often, the flanking moieties in the human flanking regions will be substituted with the corresponding moieties of the CDR donor antibody to preferably alter the binding to the antigen. These substitutions in the flanking region are identified by procedures well known in the art, for example, by modeling the interactions of the CDR residues and

from the flanking region to identify flanking residues important for antigen binding and sequence comparison to identify unusual flanking residues at particular positions. See, for example, Queen et al., U.S. Patent Nos. 5,530,101; 5,585,089; 5,693,761; 5,693,762; 6,180,370 (each of which is incorporated by reference in its entirety). Antibodies can be humanized using a variety of techniques known in the art including, for example, CDR grafting (EP 239,400; PCT Publication WO 91/09967; U.S. Patent Nos. 5,225,539; 5,530,101 and 5,585,089) , coating or coating (EP 592.106; EP 519.596; Padlan, Mol. Immunol., 28: 489 498 (1991); Studnicka et al., Prot. Eng. 7: 805 814 (1994); Roguska et al., Proc Natl. Acad. Sci. USA, 91: 969 973 (1994), and redistribution of chains (US Patent No. 5,565,332), all of which are hereby incorporated by reference in their entirety. IL-2R described herein include humanized antibodies, such as humanized mouse antibodies, fully human antibodies, and mouse antibodies.

An "anti-IL-2R antibody" is an antibody that specifically binds to an IL-2 receptor. For example, in some embodiments, an anti-IL-2R antibody binds to the high affinity IL-2 receptor (Kd ~ 10 pM). This receptor is a membrane receptor complex consisting of two subunits: IL-2R-alpha (also known as T-cell activation antigen (Tac), CD25, or p55) and IL-2R-beta (also known as p75 or CD122). In other embodiments, an anti-IL-2R antibody binds to the intermediate affinity IL-2 receptor (Kd = 100 pM), which consists of the p75 subunit and a gamma chain. In other embodiments, an anti-IL-2R antibody binds to the low affinity receptor (K d = 10 nM), which is formed only by p55.

Anti-IL-2R antibodies suitable for use in the methods described herein include monoclonal antibodies, chimeric antibodies, humanized antibodies, or fully human antibodies. Examples of anti-IL-2R antibodies capable of binding to Tac (p55) include, but are not limited to, Zenapax®, the chimeric basiliximab antibody (Simulect®), BT563 (see Baan et al., Transplant. Proc. 33: 224- 2246, 2001), and 7G8, and HuMax-TAC in development by Genmab. The mik-beta1 antibody specifically binds to the human IL-2R beta chain. Additional antibodies that specifically bind to the IL-2 receptor are known in the art. For example, see U.S. Patent No. 5,011,684; U.S. Patent No. 5,152,980; U.S. Patent No. 5,336,489; U.S. Patent No. 5,510,105; U.S. Patent No. 5,571,507; U.S. Patent No. 5,587,162; U.S. Patent No. 5,607,675; U.S. Patent No. 5,674,494; U.S. Patent No.

5,916,559.

6.2

 Detailed description

MS is an inflammatory / demyelinating CNS disease that is one of the leading causes of neurological disability in young adults (Bielekova, B. and Martin, R., 1999, Curr Treat Options Neurol. 1: 201219). The pathogenesis observed in MS patients is, at least in part, attributable to aberrant activation of T lymphocytes. Daclizumab is a humanized antibody that binds to the IL-2R alpha chain (also known as T lymphocyte activation antigen (TAC) ), CD25 or p55). CD25 is present at low levels in resting human T lymphocytes, but is significantly positively regulated in activated T lymphocytes, enabling them to receive a high affinity IL-2 signal (Waldmann, et al., 1998, Int Rev Immunol, 16: 205 -226). It has been proposed that selective binding of CD25 by daclizumab inhibits IL-2R signal transduction events (Goebel, J., et al., 2000, Transplant Immunol, 8: 153-159, Tkaczuk, J., 2001, Transplant Proc, 33: 212-213) and blocks T lymphocyte activation (Queen, C., et al., 1989, Proc Natal Acad Sci USA, 86: 10029-10033).

The procedures described herein are based on the discovery that rapid reductions in HLA-DR + CD4 + activated T lymphocytes are observed in MS patients treated with daclizumab (see FIG. 2). Rapid reductions in the amounts of activated T lymphocytes are consistent with the reductions in Gd-CELs observed in MS patients treated with daclizumab (Montalban, X. et al., Multiple Sclerosis, 13: S18-S18 Suppl. 2 OCT 2007 ; and, Kaufman, MD, et al., Neurology, 70 (11): A220-A220 Suppl. 1 MAR 11 2008). These observations suggest that changes in the level of activated HLA-DR + CD4 + T lymphocytes would be a useful tool to control the clinical response in patients with MS to daclizumab or an anti-IL-2R antibody capable of inhibiting signal transduction events. IL-2R.

Several studies have investigated the expression of HLA-DR + in blood samples obtained from patients with autoimmune diseases, including MS, to determine whether the expression of HLA-DR + could be used to control disease progression during drug therapy. None of these studies have demonstrated that HLA-DK + expression could be used to control disease progression in response to drug therapy. For example, Ferrarini et al., Controlled the expression of HLA-DR and CD25 in T lymphocytes, as well as circulating CD3 + HLA-DR + and CD4 + CD25 + cells in patients with relapsing-remitting multiple sclerosis during therapy with interferon beta 1b and determined that HLA-DR expression could not be used to control MS activity during therapy with IFN beta 1b (Ferrarini et al., 1998, Multiple Sclerosis, 4: 174-177). Other studies have also attempted to establish a link between the expression of T lymphocyte activation markers including HLA-DR and disease activity and / or drug therapy in MS (see, for example, Tang, MT, et al. , 2008, Multiple Sclerosis, 14: S181-S181 Suppl. 1; Aristimuno, C., et al., 2008, Journal Neuroimmunology, 204: 131-135; Heinrich, A., et al., 2006, Acta Neurol Scand, 113: 248-255; Sanchez-Ramon, S., et al., 2005, Immunology Letters, 96: 195-201; Gelati, M., et al., 1999, J Neurol, 246: 569-573; Genc, K., et al., 1997, J Clin Invest,

1: 2664-2671; and Bever, C., et al., 1991, Int J Immunopharmac, 13: 613-618).

Bielekova et al. Reported that in MS patients treated with daclizumab there was a gradual decrease in circulating CD4 + and CD8 + T lymphocytes and a significant expansion of bright natural cytolytic (NK) CD56 lymphocytes in vivo and this effect was highly correlated with the response to daclizumab (Bielekova, B., et al., 2006, Proc Natl Acad Sci USA, 103: 5941-5946). It was proposed that the positive correlations between the expansion of bright CD56 NK cells and the contraction of the amounts of CD4 + and CD8 + T lymphocytes support the existence of an immunoregulatory pathway in which activated CD56 NK cells inhibit T lymphocyte survival (Bielekova, B ., et al., 2006, Proc Natl Acad Sci USA, 103: 5941-5946).

Although Bielekova et al. (Bielekova, B., et al., 2006, Proc Natl Acad Sci USA, 103: 5941-5946) reported statistically significant reductions from the initial measurement in total CD4 + T cell counts (mean = -11.2%, p = 0.009) when MS patients (n = 22) were treated with daclizumab in combination with IFN beta therapy, the observation of reduced levels of CD4 + T lymphocyte counts associated with daclizumab treatment was not reproduced during the blind CHOICE study, placebo controlled. During the CHOICE study, CD4 + T lymphocyte counts were obtained as often as at 10 specific times for each subject over a course of 44 weeks of study, of approximately 65 subjects, and the statistically significant change percentages from the initial measurement (parametric analysis within the treatment groups) (p <0.05) were limited to a reduction (mean = 18.2%) in the placebo treatment group at week 2 (n = 16), and a reduction (mean = 19.1%) in the treatment group with a high dose of daclizumab at week 12 (n = 19); There were no significant changes for the low dose treatment group of daclizumab.

As shown in Table 1, the reductions observed in CD4 + T cell counts were fortuitous and showed no treatment-related trend. The P values shown in Table 1 were obtained from an ANOVA comparison with the absolute cell counts of the IFN-beta + placebo group at week 22 of the treatment phase, and therefore the changes from baseline levels in patients. CD4 + T lymphocyte counts observed in the two daclizumab treatment groups of the CHOICE study were not statistically significant compared to the placebo group and were considered fortuitous. The number of patients sampled within each treatment group is shown in parentheses (n).

Table 1. Changes associated with daclizumab in CD4 + T cell counts.

Immune subset

Treatment Basal cell average / mm3 Average of cells in week 22 / mm3 Average exchange rate at week 22 P value at week 22 (n)

CD4 + T lymphocytes

IFN-beta + placebo 735 606 -5.2 (13)

IFN-beta + DAC 1 mg / kg

789 697 -16.8 0.74 (18)

IFN-beta + DAC 2 mg / kg

652 546 -20.9 0.82 (19)

CD4 + HLA-DR +

IFN-beta + placebo 31.2 32.2 -7.3 (13)

IFN-beta + DAC 1 mg / kg

31.1 20.2 -45.7 0.006 (18)

IFN-beta + DAC 2 mg / kg

27.6 20.3 -32.7 0.009 (17)

In addition, the subset of HLA-DR + CD4 + T lymphocytes typically represents a small percentage of all CD4 + T lymphocytes present in both healthy human subjects and diseased human subjects. The average percentage range of HLA-DR + CD4 + T lymphocytes for placebo subjects at all point-controlled times during the CHOICE study was 5.5% to 6.3% of all CD4 + T lymphocytes. The maximum mean reductions from the initial measurement in HLA-DR + CD4 + T lymphocyte counts determined during the CHOICE study, the maximum average reduction in the low dose group of daclizumab = -38.9%, the maximum average reduction in the group High-dose daclizumab = -48.4%, were much greater than the -11.2% change in total CD4 + T lymphocytes presented by Bielekova et al. (Bielekova, B., et al., 2006, Proc Natl Acad Sci USA, 103: 59415946). The magnitude of the change observed in HLA-DR + CD4 + T lymphocyte counts in response to daclizumab treatment in the CHOICE study, supports the use of HLA-DR + CD4 + T lymphocyte counts as a sensitive marker to control the efficacy of anti-IL-2R antibody therapy in MS patients.

This document describes procedures for the use of HLA-DR + CD4 + T cells as a marker for the activity of anti-IL2R antibodies in MS patients. Typically, blood samples from patients with MS are analyzed for cell surface markers using flow cytometry (see, for example, Bielekova, B., et al., 2006, Proc Natl Acad Sci USA, 103: 5941-5946) . For example, the amount of HLA-DR + CD4 + T lymphocytes can be analyzed using commercially available antibodies that preferentially bind to the HLA-DR + protein, and in combination with fluorescent activated cell sorting (FACS), determine the levels of cells that express HLA-DR +. The percentage change in the amount of HLA-DR + CD4 + T cells in response to treatment can be used to control the efficacy of an anti-IL-2R antibody.

In some embodiments, a reduction in the amount of HLA-DR + CD4 + T lymphocytes is observed in response to treatment with an antibody against IL-2R. The amount of HLA-DR + CD4 + T lymphocytes can be determined by absolute cell count, that is, the number of cells per mm3 or mm2, or as a percentage of total CD4 + T lymphocytes. Depending on the individual patient and the relapsing form of MS, the overall reduction in the amount of HLA-DR + CD4 + T cells may vary from 25% to 100%. For example, in some embodiments, the amount of HLA-DR + CD4 + T lymphocytes can be reduced by at least 25%, at least 30%, at least 35%, at least 40%, at least one 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80 %, at least 85%, at least 90%, at least 95%, or at least 100%.

In some embodiments, additional cell surface markers can be monitored to provide additional information regarding the clinical response in MS patients treated with an anti-IL-2R antibody. Additional cell surface markers include, but are not limited to, CD3, bright CD56 NK cells, CD4, CD25, CD16, CD122, and CD8. Tests for the determination of these markers have been described, see, for example, Bielekova, B., et al., 2006, Proc Natl Acad Sci USA, 103: 5941-5946. For example, in some embodiments, the amount of HLA-DR + CD4 + T lymphocytes and the amount of bright CD56 NK cells can be analyzed and used to control the efficacy of an anti-IL-2R antibody.

The determination of the amount of HLA-DR + CD4 + T lymphocytes generally requires taking more than one sample from a patient at selected times. The determination of the sampling time is not critical to the procedures described herein and can be selected by a physician based, in part, on the amount of time a patient has been treated with an anti-IL-2R antibody. Other factors that may affect the time of sampling include, but are not limited to, the amount of time the patient has been treated for MS, the therapy or therapies the patient has received before treatment with an anti-IL-2R antibody, and if the patient is showing one or more of the following symptoms: increased recurrence rate, an increase in the value of the Expanded Disability Level Scale (EEND), an increased amount of new or increased Gd-CEL, and an increase in new or increased MRI T2 lesions. See, for example, Perini et al., 2004, J Neurology, 251: 305309; Sorensen, et al., 2003 Lancet, 362: 184-191; Pachner, 2003, Neurology, 61 (Suppl. 5): S2-S5; Perini et al., 2004, J Neurology, 251: 305-309; and Farrell, et al., 2008, Multiple Sclerosis, 14: 212-218.

For example, in some embodiments, changes in the amount of HLA-DR + CD4 + T cells can be monitored before, during and after the start of treatment with an anti-IL-2R antibody.

By way of another example, changes in the amount of HLA-DR + CD4 + T lymphocytes can be monitored before the start of treatment with an anti-IL-2R antibody and at intervals selected thereafter (for example, weekly, monthly, once every two months, once every three months, once every six months).

As another example, changes in the amount of HLA-DR + CD4 + T lymphocytes can be monitored after the start of treatment (for example, in a week, in a month, in two months, in three months, in six months) with an anti-IL-2R antibody and at selected intervals thereafter (for example, weekly, monthly, once every two months, once every three months, once every six months).

Typically, a 25% reduction or greater in the amount of HLA-DR + CD4 + T lymphocytes is consistent with the administration of a therapeutically effective dose of an anti-IL-2R antibody. As used herein, a "therapeutically effective dose" is a dose sufficient to prevent progression, decrease the recurrence rate, or reduce one or more of the symptoms associated with the progression of the disease in multiple sclerosis.

For example, in some embodiments, the administration of a therapeutically effective dose of an antiIL-2R antibody to an MS patient decreases the amount of recurrences by at least one that occurs in a given period of time, such as 1 year, in the treated patient Recurrences are typically assessed by history and physical examination defined as the appearance of a new symptom or the worsening of an old symptom attributable to multiple sclerosis, accompanied by a new appropriate neurological abnormality or focal neurological dysfunction that lasts at least 24 hours in absence of fever, and preceded by stability or improvement for at least 30 days (see, for example, Sorensen, et al., 2003 Lancet, 362: 1184-1191).

In other embodiments, the administration of a therapeutically effective dose of an anti-IL-2R antibody to an MS patient decreases the amount of lesions detected in the patient's brain. Magnetic resonance imaging (MRI) of the brain is an important tool for understanding the dynamic pathology of

multiple sclerosis T2-weighted brain MRI defines lesions with high sensitivity in multiple sclerosis and is used as a measure of disease burden. However, such high sensitivity occurs at the expense of specificity, since changes in the T2 signal may reflect areas of edema, demyelination, gliosis and axonal loss. It is believed that the areas of Gd-CEL demonstrated in MRI of T1-weighted brain reflect underlying alteration of the brain-stem barrier from active perivascular inflammation. These areas of potentiation are transient, typically lasting <1 month. MRI of the brain due to Gd-CEL is therefore used to assess disease activity. The majority of T2-weighted lesions (T2) in the central white matter of subjects with multiple sclerosis begins with a variable period of Gd-CEL. Gd-CEL and T2 lesions represent phases of a single pathological process. MRI of the brain is a conventional technique to assess Gd-CEL and T2 lesions and is routinely used to assess disease progression in MS (for example, see Lee et al., Brain 122 (Pt 7): 1211 -2, 1999).

As shown in FIG. 1, the treatment of MS patients with daclizumab reduced the average amount of new and increased Gd-CEL by 25% or more. Therefore, in some embodiments a therapeutically effective dose of an anti-IL-2R antibody for an MS patient decreases the amount of Gd-CEL detected in the patient's brain by approximately 25% to 80%. In some embodiments, the amount of Gd-CEL detected in the patient's brain is decreased by at least 25%, by at least 30%, by at least 35%, by at least 40%, by at least one 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80 %, at least 85%, at least 90%, at least 95%, and at least 100%.

In other embodiments, administration of a therapeutically effective dose of an anti-IL-2R antibody to an MS patient decreases the amount of T2 MRI lesions detected in the patient's brain.

In other embodiments, the administration of a therapeutically effective dose of an anti-IL-2R antibody to an MS patient stabilizes the progression of a patient's disability determined by the "Expanded Disability Level Scale (EEND)" that can be used to Classify neurological abnormality in MS patients (Kurtzke, 1983, Neurology, 33-1444-52). The EEND comprises 20 degrees from 0 (normal) to 10 (death due to MS), which progress in a single-point stage from 0 to 1 and in stages of 0.5 ascending points. The values are based on a combination of functional values of the system, the degree of mobility of the patient, the need for walking assistance, or assistance in activities of daily living. The functional values of the system measure the function within individual neurological systems including visual, pyramidal, cerebellar, brainstem, sensory, intestinal and bladder, cerebral and other functions.

In other embodiments, administration of a therapeutically effective dose of an anti-IL-2R antibody to an IFN-beta positive MS NAb patient reduces a patient's disability value by 10% to 75%. For example, in some embodiments, a disability value of the patient can be reduced by at least 10%, by at least 15%, by at least 20%, by at least 25%, by at least 30%, by at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at at least 70%, or at least 75%.

In some embodiments, no changes or an increase in the amount of HLA-DR + CD4 + T lymphocytes will be observed after treatment of an MS patient with an anti-IL-2R antibody. In these embodiments, patients can be evaluated to determine if they are responding poorly or are not able to respond to treatment with an anti-IL-2R antibody. MS patients who are responding poorly to therapy generally have a higher average recurrence rate, a higher risk of experiencing a second recurrence, a higher risk of having a sustained progression of> 1 over the END, and a lower probability of being relapse-free (Malucchi, et al., 2004, Neurology, 62: 2031-2037). Accordingly, several clinical assessment criteria can be used to determine if a patient is responding to treatment with an anti-IL-2R antibody including the frequency and recurrence rate, an increase of 1 point or more in the value of the Expanded Scale of Level of Disability (EEND), an increase in the amount of Gd-CEL, and / or an increase in the amount of MRI T2 lesions.

The data required to determine the clinical assessment criteria can be collected at the start of anti-IL-2R treatment and / or during follow-up visits. In some embodiments, MS patients who are responding poorly to an anti-IL-2R antibody can be treated with additional agents. For example, in some embodiments, one or more anti-IL-2R antibodies can be administered to an MS patient. In other embodiments, an anti-IL-2R antibody may be administered in combination with another MS therapy, such as an IFNbeta product. Examples of suitable IFN-beta products include, but are not limited to, one of the three IFN-beta products that have been approved: IFN-beta-1b (Betaferon®, Schering AG, Berlin, Germany), IFN-beta-1a (Avonex ®, Biogen Idec, Cambridge MA), and IFN-beta-1a (Rebif®, Ares-Serono, Geneva, Switzerland). Non-limiting examples of other commercialized drugs that may be used in combination with an anti-IL-2R treatment include glatiramer acetate (e.g., Copaxone®, Teva Pharmaceutical Industries, Ltd., Israel), corticosteroids, riluzole, azathioprine, cyclophosphamide, methotrexate. , and mitoxantrone.

MS patients suitable for treatment with an anti-IL-2R antibody have typically been diagnosed with a relapsing form of multiple sclerosis including relapsing-remitting multiple sclerosis, secondary progressive multiple sclerosis, progressive relapsing multiple sclerosis and aggravating recurrent multiple sclerosis.

By "relapsing-remitting multiple sclerosis" in this document is meant a clinical course of MS that is characterized by clearly defined acute attacks with complete or partial recovery and without disease progression between attacks. By "secondary progressive multiple sclerosis" in this document is meant a clinical course of MS that is initially relapsing-remitting, and then becomes progressive at a variable rate, possibly with occasional recurrence and minority remission. The term "progressive relapsing multiple sclerosis" means a clinical course of MS that is progressive from the onset, alternated by recurrences. There is a significant recovery immediately after a recurrence, but between recurrences there is a gradual worsening of disease progression. The term "aggravated recurrent multiple sclerosis" means a clinical course of MS with unpredictable recurrences of symptoms, from which people do not return to normal and do not recover completely.

In some embodiments, the anti-IL-2 receptor antibody is daclizumab. Recombinant genes encoding daclizumab are a compound of human (approximately 90%) and murine (approximately 10%) antibody sequences. The donor murine anti-Tac antibody is an IgG2a monoclonal antibody that specifically binds to the IL-2R Tac protein and inhibits the IL-2-mediated biological responses of lymphoid cells. The murine anti-Tac antibody was "humanized" by combining the complementarity determining regions and other selected moieties of the murine anti-TAC antibody with the flanking and constant regions of the human IgG1 antibody. The humanized anti-Tac daclizumab antibody is described and its sequence is set forth in U.S. Patent No. 5,530,101, see SEQ ID No. 5 and SEQ ID No. 7 for the heavy and light chain variable regions respectively. SEQ ID Nos. 5 and 7 of United States Patent No.

5,530,101 are disclosed as SEQ ID NO. 1 and 2 respectively in the sequence listing presented therewith. U.S. Patent No. 5,530,101 and Queen et al., Proc. Natl. Acad. Sci. 86: 1029-1033, 1989 both are incorporated by reference herein in their entirety.

Daclizumab has been approved by the US Food and Drug Administration (FDA) for the prophylaxis of acute organ rejection in subjects receiving kidney transplants, as part of an immunosuppressive regimen that includes cyclosporine and corticosteroids and is marketed by Roche as ZENAPAX®. Daclizumab has also been shown to be active in the treatment of tropical spastic myelopathy / paraparesis associated with type 1 human T lymphocyte lymphotrophic virus (HAM / TSP, see Lehky et al., Ann. Neuro., 44: 942-947, 1998) . The use of daclizumab to treat posterior uveitis has also been described (see Nussenblatt et al., Proc. Natl. Acad. Sci., 96: 7462-7466, 1999).

Antibodies that bind to the same epitope (or overlap) as daclizumab can be used in the methods disclosed herein. In some embodiments, the antibody will have at least 90%, at least 95%, at least 98%, or at least 99% sequence identity with daclizumab. The antibody can be of any isotype, including but not limited to IgG1, IgG2, IgG3 and IgG4.

In some embodiments, the antibody is basiliximab, marketed as Simulect® by Novartis Pharma AG. Basiliximab is a chimeric antibody (murine / human), produced by recombinant DNA technology that functions as an immunosuppressive agent, specifically binding to and blocking the alpha chain of IL-2R on the surface of activated T lymphocytes.

Anti-IL-2R antibodies can be administered parenterally, that is, subcutaneously, intramuscularly, intravenously, intranasally, transdermally, or by a needleless injection device. Compositions for parenteral administration will usually include a solution of an anti-IL-2R antibody in a pharmaceutically acceptable carrier. Pharmaceutically acceptable non-toxic carriers or diluents are defined as vehicles commonly used to formulate pharmaceutical compositions for administration to animals or humans. See, for example, Remington: The Science and Practice of Pharmacy, A.R. Gennaro, 20th Edition, 2001, Lippincott Williams & Wilkins, Baltimore, MD, for a description of compositions and formulations suitable for pharmaceutical delivery of the anti-IL-2R antibodies disclosed herein. See US Patent Application Publications No. 2003/0138417 and 2006/0029599 for a description of liquid and lyophilized formulations suitable for the pharmaceutical supply of daclizumab.

Procedures for preparing pharmaceutical compositions are known to those skilled in the art (see Remington: The Science and Practice of Pharmacy, supra). In addition, the pharmaceutical composition or formulation may include other vehicles, adjuvants, or non-toxic, non-therapeutic and non-immunogenic stabilizers and the like. The effective amounts of said diluent or carriers will be those amounts that are effective in obtaining a pharmaceutically acceptable formulation in terms of solubility of components, or biological activity.

The concentration of antibody in the formulations can vary widely, that is, from less than about 0.5%, usually at or at least about 1% to as much as 15 or 20% by weight or 1 mg / ml. at 100 mg / ml. The concentration is selected primarily based on fluid volumes, viscosities, etc., according to the particular mode of administration selected.

Generally, an adequate therapeutic dose of daclizumab is from about 0.5 milligrams per kilogram (mg / kg) to about 5 mg / kg, such as a dose of about 0.5 mg / kg, about 1 mg / kg, of approximately 1.5 mg / kg, approximately 2 mg / kg, approximately 2.5 mg / kg, approximately 3.0 mg / kg, approximately 3.5 mg / kg, approximately 4.0 mg / kg , about 4.5 mg / kg, or about 5.0 mg / kg administered intravenously or subcutaneously. Unit dosage forms are also possible, for example 50 mg, 100 mg, 150 mg, 200 mg, 300 mg, 400 mg, or up to 500 mg per

5 doses

Other dosages may be used to obtain serum levels of 2 to 5 μg / ml that are necessary for saturation of the Tac subunit of the IL-2 receptor to block activated T cell responses. Higher levels such as approximately 5 to 40 μg / ml may be necessary for clinical efficacy. A specialist in the art will be able to build an administration regimen to maintain serum levels within the range.

10 from 2 to 40 μg / ml.

Basiliximab doses are likely to be lower, for example from 0.25 mg / kg to 1 mg / kg, for example, 0.5 mg / kg, or unit doses of 10, 20, 40, 50 or 100 mg, due to the higher affinity of basiliximab for the IL-2R target. The general principle to keep the IL-2 receptor saturated can be used to guide the choice of dose levels of other antibodies against IL-2R.

15 Single or multiple administrations of anti-IL-2R antibodies can be made with dosages and administration frequencies selected by the physician. Generally, multiple doses are administered. For example, multiple administration of daclizumab or other anti-IL-2R antibodies may be used, such as monthly, bi-monthly administration, every 6 weeks, every other week, weekly or twice a week.

7. Examples

20 Example 1: CHOICE study

The CHOICE study was a multicenter, placebo-controlled, double-blind, randomized, phase 2 study of subcutaneous daclizumab (SC) added to interferon (IFN) -beta in the treatment of active, relapsing forms of MS. The results of the CHOICE study confirmed that daclizumab at 2 mg / kg every two weeks significantly decreased the amount of new Gd-CEL in patients who have active, relapsing forms of MS in

25 concurrent therapy with IFN-beta (Montalban, X. et al., Multiple Sclerosis, 13: S18-S18 Suppl. 2 OCT 2007; and, Kaufman, MD, et al., Neurology, 70 (11): A220-A220 Suppl. 1 MAR 11 2008). A minor decrease in new or increased Gd-CEL was observed for those study subjects receiving 1 mg / kg of daclizumab every four weeks.

One patient who was randomized was included in the study. The included patients remained in their

30 baseline IFN-beta regime and were randomized in a 1: 1: 1 ratio to one of the following 3 treatment arms (see Table 2).

Table 2

Treatment arm1 Dose level and frequency No. of total visits of No. of patients dosing

A (High dose) 2 Daclizumab SC: 2 mg / kg q2 weeks 11 55 x 11 doses

B (Low dose) 3 Daclizumab SC: 1 mg / kg q4 weeks 11 55 x 6 doses

C (Placebo) 4 Placebo SC: q2 weeks x 11 doses 11 55

1 All patients continue with the previous regimen of IFN-beta SC / IM for the duration of the study. 2 Patients in arm A (high dose) receive 2 SC injections (2 daclizumab 1 mg / kg) for 11 Dosing visits Maximum dose of daclizumab per dosing visit = 200 mg. 3 Patients in arm B (low dose) receive 2 SC injections (1 daclizumab 1 mg / kg, 1 placebo) during 6 dosing visits, alternating with 2 SC injections (2 of placebo) during 5 dosing visits. Maximum dose of daclizumab per dose visit of daclizumab = 100 mg. 4 Patients in arm C (placebo) receive 2 SC injections (2 placebo) during 11 dosing visits.

The selection period was up to 3 weeks. The treatment period was designated as 24 weeks (6 months, until day 168) to include 4 weeks after the last dose of the blind study drug (Dose # 11, which happens on visit # 14, day 140). After the treatment period, the 35 patients were followed for a total of 48 weeks (12 months) and IFN beta therapy was continued for at least 5 months of this period. The maximum total time in the study for each patient was approximately 18 months.

Evaluations of a patient given by the EEND and the Composite Functional Scale of Multiple Sclerosis, version 3 (MSFC-3) were performed by a physician who was not involved in the treatment of the patient and was appointed

an "evaluation doctor". All other assessments of the patient were under the responsibility of the physician in charge of the patient's treatment (treating physician). The MSFC-3 includes quantitative trials of: (1) Leg function / locomotion-25-step timed walk (T25FW); (2) Function of the arms-9 hole stakes test (9HPT), and (3) Cognition test-Auditory addition in serial rhythm with intervals between stimuli of 3 seconds (PASAT3) (Cutter et al., 1999, Brain , 122 (Pt 5): 871-882).

Preliminary eligibility for the CHOICE study was established by history, inspection of charts, and routine evaluations. During the treatment and follow-up period, several procedures and evaluations were performed on the subjects on specified days including, but not limited to, RM, EEND, MSFC-3, physical examinations, physical examinations aimed at symptoms, hematology / serum chemistry (by example, for the determination of pharmacokinetic evaluation and anti-DAC antibodies), and blood extractions for pharmacodynamic evaluations and IFN-beta NAb.

The drug substance daclizumab manufactured by PDL BioPharma, Inc. (Redwood City, CA) for subcutaneous delivery, was supplied in single-use vials containing 100 mg daclizumab in 1.0 ml of 40 mM sodium succinate, 100 sodium chloride mM, 0.03% polysorbate 80, pH 6.0. The placebo was given in single-use vials as an isotonic solution in matching vials containing 40 mM sodium succinate, 6% sucrose, 0.03% polysorbate 80, pH 6.0.

Example 2: Reductions in HLA-DR + CD4 + T lymphocytes in response to treatment with daclizumab

Changes in the amount of HLA-DR + CD4 + T lymphocytes were determined in patients participating in the CHOICE study (see Example 1). HLA-DR + CD4 + T lymphocyte levels were analyzed in patients who received daclizumab 2 mg / kg every 2 weeks (high dose), daclizumab 1 mg / kg every 4 weeks (low dose), or placebo SC added to therapy with Interferon beta for 24 weeks. Samples of whole blood were collected from patients in vacutainer tubes containing anti-coagulant using conventional venipuncture techniques and sent by overnight courier for immediate analysis, blind to treatment using fluorescent activated cell sorting (FACS). Commercially available antibodies were obtained from BD Biosciences (USA) that preferentially bind to the HLA-DR protein (e.g., the L243 antibody clone (G45-6)) and that preferentially bind to specific immune subsets (e.g. , antibodies that selectively bind to the CD3 (for example, clone SK7) and CD4 (for example, clone SK3) antigens and were used in combination with FACS to determine the levels of cells expressing HLA-DR + when patients they were treated with antibody against DAC. Sample preparation and FACS test procedures were performed in accordance with the practices recommended in the "Guideline for Flow Cytometric Immunophenotyping" (see Calevetti et al., 1993, Cytometry 14: 702-714). Parametric (t-test for dependent samples) and non-parametric (Wilcoxon) tests comparing the levels of cellular counts of immune subsets between dosage groups were performed. The individual characteristics of daclizumab exposure (results of post hoc analysis) of a subset of subjects, including the steady state sine (Css, min), and the AUCss, separately, were used as predictors to model changes from baseline in individual immune subsets or changes in time (area calculated under the change curve from the initial measurement-time (AUC)). The relationship between changes from the basal level of putative subsets of activated T lymphocytes and the total of new or increased Gd-CELs between weeks 8 and 24 was also evaluated using statistical analysis approximations of linear correlation, negative binomial correlation, ANOVA or Kruskal-Wallis essays.

Analysis of daclizumab treatments at a dose of 2 mg / kg SC every 2 weeks, added to the background treatment with interferon-beta produced statistically and clinically significant reductions in the activity of MS lesions (see FIG. 1) in comparison with the rest with only interferon-beta (and adding the placebo).

Rapid reductions in absolute levels of activated T lymphocytes were observed between patients in both dose groups of daclizumab, but not in patients treated with placebo (see FIG. 2). As shown in FIG. 2, decreases were observed from the initial measurement in the absolute levels of CD4 + T lymphocytes activated HLA-DR + for both dosage groups. Statistically significant reductions are indicated (p <0.05) compared to placebo (* low dose DAC and ** high dose DAC). The absolute levels of activated CD4 + T lymphocytes HLA-DR + returned to approximate baseline levels at the time of de-saturation of CD25 between days 196 and 280. In the last sampling at the end of the treatment period (week 22), the mean values in HLA-DR + CD4 + T lymphocytes they were significantly lower in both DAC dose groups compared to placebo (low dose DAC versus placebo: 22.5 versus 37.4 cells / ,l, P = 0.006; DAC high dose versus placebo: 23.1 versus 37.4 cells / ,l, P = 0.009). Reductions in HLA-DR + CD4 + T cells reversed after discontinuation of DAC treatment.

As shown in FIG. 3, a statistically significant positive linear relationship (p = 0.008) was observed between individual exposure to steady-state DAC and the subject's reductions from the initial measurement in absolute levels of HLA-DR + CD4 + T lymphocyte counts over time (within the dosing period), when each factor was expressed in terms of area under the curve (AUC) and when a reduction in HLA-DR + CD4 + T lymphocyte counts was expressed as a positive value (i.e. when a decrease in HLA-DR + CD4 + T lymphocyte counts from the initial measurement it was represented as a positive value). Therefore, a significant positive relationship between exposure to daclizumab and the reduction in blood count was demonstrated.

T lymphocytes HLA-DR + CD4 + T (AUC) over time.

As shown in FIG. 2 and 3, the treatment of patients with MS with daclizumab was associated with reductions in HLA-DR + CD4 + T lymphocytes; these reductions were rapid during the daclizumab treatment phase for both dosage groups and reversed after the end of daclizumab treatment (see FIG. 2). A positive, dose-dependent and statistically significant relationship was observed between exposure to daclizumab (AUCss) and reductions in HLA-DR + CD4 + T cell counts (AUC) over time when a reduction in T lymphocyte counts HLA-DR + CD4 + was expressed as a positive value (i.e. when a decrease in T lymphocyte counts HLA-DR + CD4 + from the initial measurement was expressed as a positive value) (see FIG. 3). The effect dependent on the observed exposure of daclizumab in the reduction of

10 amounts of activated T lymphocytes during the treatment phase is consistent with the exposure-dependent clinical response. Taken together, these observations support the use of HLA-DR + CD4 + T lymphocyte count control as a biomarker of daclizumab antibody treatment in MS.

Although various specific embodiments have been illustrated and described, it will be appreciated that various changes can be made without departing from the scope of the invention.

15 LIST OF SEQUENCES

<110> Facet Biotech corporation

<120> PROCEDURES TO CONTROL THE EFFECTIVENESS OF ANTI-IL-2R ANTIBODIES IN 20 PATIENTS WITH MULTIPLE SCLEROSIS

<130> 222 WO 01

<150> US 61 / 173,138 25 <151>

<160> 2

<170> PatentIn version 3.5

30 <210> 1

<211> 116

<212> PRT

<213> Artificial

35 <220>

<223> Synthetic construction. "Variable region of the heavy chain of the humanized anti-Tac PDL antibody"

<400> 1 40

<210> 2

<213> Artificial

<220>

<223> Synthetic construction. "Variable region of the 10 PDL anti-Tac humanized antibody light chain"

<400> 2

Claims (15)

one.

 A method of monitoring the efficacy of an anti-IL-2R antibody in a patient diagnosed with multiple sclerosis, which comprises determining the level of HLA-DR + CD4 + T lymphocytes, in which a decrease in the level of HLA- T lymphocytes DR + CD4 + in the patient after exposure to the anti-IL-2R antibody indicates that the anti-IL-2R antibody is effective in improving at least one symptom of multiple sclerosis in the treated patient.

2.

 The method according to claim 1, wherein the level of HLA-DR + CD4 + T lymphocytes is determined in blood samples from a patient before, and subsequently, administering the anti-IL-2R antibody to said patient, in which a decrease in those in HLA-DR + CD4 + T lymphocytes after said treatment indicates that the anti-IL-2R antibody is effective in improving at least one symptom of multiple sclerosis in the treated patient.

3.

 A method according to claim 1, wherein the method comprises comparing the amount of HLA-DR + CD4 + T lymphocytes in a blood sample obtained from the patient with an untreated reference, in which a decrease in the amount of lymphocytes T HLA-DR + CD4 + in the blood sample compared to the untreated reference indicates that the treatment is effective in improving at least one symptom of multiple sclerosis in the subject.

Four.

 The method according to claim 1, 2 or 3, wherein the antibody that specifically binds to the interleukin 2 receptor is a humanized antibody.

5.

 The method according to claim 1, 3 or 4, wherein the anti-IL-2R antibody specifically binds to the alpha subunit of the high affinity human receptor of interleukin-2 and inhibits IL-2 signaling.

6.

 The method of claim 5, wherein the anti-IL-2R antibody is a humanized antibody.

7.

 The method of claim 6, wherein the humanized antibody is daclizumab.

8.

The method according to any one of the preceding claims wherein the improvement of a multiple sclerosis symptom comprises reducing the amount of recurrences in a given period, reducing the rate of increase of the value on the expanded scale of the subject's level of disability. , reduce the amount of MRI lesions enhanced with gadolinium T1 contrast, or reduce the amount of MRI T2 lesions.

9.

 The method of claim 7, wherein daclizumab is administered at a dose of about 0.5 to about 5 milligrams per kilogram, or at a dose of about 1 to about 2 milligrams per kilogram.

10.

 The method of claim 7, wherein daclizumab is administered intravenously, subcutaneously, intramuscularly, intranasally, or transdermally.

eleven.

 The method of claim 7, wherein daclizumab is administered at least biweekly, or at least monthly.

12.

 The method according to any one of the preceding claims, wherein the subject has a relapsing form of multiple sclerosis.

13.

 The method according to claim 12, wherein the subject has relapsing relapsing, secondary progressive, progressive recurrent, or aggravated recurrent multiple sclerosis.

14.

 A method according to claim 1, comprising determining the amount of HLA-DR + CD4 + T lymphocytes in blood samples collected from said patients before, and after, administering the antiIL-2R antibody to said patient, to determine if A change in the amount of HLA-DR + CD4 + T lymphocytes occurred in the patient after treatment with the anti-IL-2R antibody.

fifteen.

 The method according to claim 14, wherein the change in the amount of HLA-DR + CD4 + T lymphocytes in the treated patient is reduced by at least 25%, at least 50% or at least 100%.